Study On Regulation Of Acupuncture On Mice Adaptive Immunity In Different States Based On Tissue-resident Memory T Cells

OBJECTTo clarify the role of tissue-resident memory T cells in acupoint localization in acupuncture influence adaptive immunity at different statesMETHODS(1)To observe the effect of acupuncture on tissue-resident memory T cells in acupoints under physiological conditions,the male C57BL/6 mice(20±2 g)were placed in isolation units for 7 days of domestication.They were randomly divided into control group(n=6)and acupuncture group(n=6).the acupuncture intervention was performed at ZT12(ZT0:7:00)at“zusanli”acupoint on the right side,once a day for10 days,30 minutes each time,and insertion was performed every 5 minutes during the acupuncture process for 1 min with a frequency of 120 times/min.The control group were bundled at the same time.After the acupuncture intervention,the skin of the“zusanli”acupoint and inguinal lymph node were taken.The number of tissue-resident memory T cells at skin tissue and memory T cell subsets in lymph node by flow cytometry.(2)To observe the effect of acupuncture on tissue-resident memory T cells in acupoints under acute inflammation.Domesticated male C57BL/6mice(20±2 g)were randomly divided into control group(n=6),model group(n=6)and treatment group(n=12)according to pain threshold.At the first phase,the treatment group was treated with acupuncture intervention once a day for 7 days,30minutes every time.During the acupuncture process,the insertion was performed for1 minute every 5 minutes,the frequency was 120 times/min.The control group and the model group mice were bundled at the same time.After the first phase of intervention,the model group and the treatment group were injected intradermally with 0.02 ml of complete Freund's adjuvant into the right footpad,and the control group was injected with the same amount of saline at the same place.The pain threshold and foot volume of the mice were measured 24 hours after modeling and the paw swelling degree was calculated.After successful modeling,the treatment group was randomly divided into the pre-acupuncture group and the model-acupuncture group,and the pre-acupuncture group no longer performed acupuncture intervention.The model-acupuncture group was treated 3 times in the same way as in phase one,and the remaining groups were bundled at the same time.After all treatment was done,the skin of the"Zusanli"area on the right side of the mouse,the skin from the affected parts and the right inguinal lymph node were extracted,and the number of memory T cell subsets was detected with flow cytometry.Results1.Under physiological conditions,acupuncture can increase the number of TRM cells in the local skin of the acupoints.The difference of TRM cells in the acupuncture group compared with the control group was statistically significant,P=0.00<0.01.Acupuncture had no significant effect on the number of TRM(P=0.163>0.05),TEM(P=782>0.05)and TCM(P=0.566>0.05)in the inguinal lymph nodes.2.In the state of acute inflammation,the number of TRM cells in the local acupoints was not significantly changed from that in the control group,P=0.104>0.05,and the number of TRM cells decreased in the inguinal lymph nodes,P=0.024<0.05,the number of TCM cells decreased,P=0.005<0.01,and the number of TEM cells increased significantly P=0.003<0.01.Pre-acupuncture can reduce the number of TRM cells in the local skin of the acupoints,and increase the number of TRM cells,the ratio of the two is significantly decreased,P=0.019<0.05.At the same time,compared with the model group,the number of CD69~-CD103~+TRM in the inguinal lymph nodes decreased significantly,P=0.042<0.05.Compared with the model group,the model-acupuncture group increased the number of CD69~-CD103~+TRM cells in the local acupoints significantly,P=0.007<0.01,and decreased the number of CD69~-CD103~+TRM cells in the inguinal lymph nodes,P=0.001<0.01.At the same time,compared with the model group,the number of TRM cells in the affected area was significantly increased,P=0.026<0.05.ConclusionAcupuncture has different effects on the regulation of tissue-resident memory T cells in acupoints under different states.In physiological conditions,acupuncture can mobilize local skin TRM cells in acupoints,thereby enhancing the body's adaptive immune response and adjusting immune function.In the acute inflammatory state,acupuncture can promote TEM activation in lymph nodes,and promote the localized tissue-resident memory T cells in the acupoints to recruit to the lesions,participate in the immune response,and produce anti-inflammatory effects.